Table 2.
Parameters used in eight trials for the robustness study *.
| Measurement Level (High–Low) | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Factor | Standard Value | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| Annealing temperature (°C) | 59 | 60 | 60 | 60 | 60 | 58 | 58 | 58 | 58 |
| 260/230 ratio | ≈2.0–2.2 | 2.1 | 2.1 | 0.7 | 0.7 | 2.1 | 2.1 | 0.7 | 0.7 |
| 260/280 ratio | ≈1.8–2.0 | 2.0 | 1.7 | 2.0 | 1.7 | 2.0 | 1.7 | 2.0 | 1.7 |
| Annealing time (s) | 30 | 40 | 40 | 20 | 20 | 20 | 20 | 40 | 40 |
| Primer concentration (ng/μL) | 10 | 15 | 5 | 15 | 5 | 5 | 15 | 5 | 15 |
| Reaction volume (μL) | 8 | 9 | 7 | 7 | 9 | 9 | 7 | 7 | 9 |
| Reagent (HRM Kit) | - | S | Q | Q | S | Q | S | S | Q |
HRM (High Resolution Melting) Kit: S (SensiFast™ HRM Kit, Meridian Biociences®) and Q (qPCRBIO™ HRM Mix, PCRBiosystems). * Based on Youden and Steiner (1975) [60].